title: Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae.
abstract: We purified a novel actin filament (F-actin)-binding protein from the soluble fraction of Saccharomyces cerevisiae by successive column chromatographies by use of the 125I-labeled F-actin blot overlay method. The purified protein showed a minimum Mr of about 140 kDa on SDS-polyacrylamide gel electrophoresis and we named it ABP140. A search with the partial amino acid sequences of ABP140 against the Saccharomyces Genome Database revealed that the open reading frame of the ABP140 gene (ABP140) corresponded to YOR239W fused with YOR240W by the +1 translational frame shift. The encoded protein consisted of 628 amino acids with a calculated Mr of 71,484. The recombinant protein interacted with F-actin and showed the activity to crosslink F-actin into a bundle. Indirect immunofluorescence study demonstrated that ABP140 was colocalized with both cortical actin patches and cytoplasmic actin cables in intact cells. However, elimination of ABP140 by gene disruption did not show a deleterious effect on cell growth or affect the organization of F-actin. These results indicate that ABP140 is not required for cell growth but may be involved in the reorganization of F-actin in the budding yeast.
authors.affiliation: Department of Molecular Biology and Biochemistry, Osaka University Medical School, Suita, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Asakura
authors.forename: T
authors.initials: T
authors.lastname: Sasaki
authors.forename: F
authors.initials: F
authors.lastname: Nagano
authors.forename: A
authors.initials: A
authors.lastname: Satoh
authors.forename: H
authors.initials: H
authors.lastname: Obaishi
authors.forename: H
authors.initials: H
authors.lastname: Nishioka
authors.forename: H
authors.initials: H
authors.lastname: Imamura
authors.forename: K
authors.initials: K
authors.lastname: Hotta
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
authors.forename: H
authors.initials: H
authors.lastname: Nakanishi
authors.forename: Y
authors.initials: Y
authors.lastname: Takai
mesh_terms: D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D003600:Cytosol / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins
chemical_list: C110542:ABP140 protein, S cerevisiae; D000199:Actins; D008840:Microfilament Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins